Neurodevelopmental assessment of normocephalic children born to Zika virus exposed and unexposed pregnant people.

BACKGROUND: Studies examining the association between in utero Zika virus (ZIKV) exposure and child neurodevelopmental outcomes have produced varied results. METHODS: We aimed to assess neurodevelopmental outcomes among normocephalic children born from pregnant people enrolled in the Zika in Pregnancy in Honduras (ZIPH) cohort study, July-December 2016. Enrollment occurred during the first prenatal visit. Exposure was defined as prenatal ZIKV IgM and/or ZIKV RNA result at enrollment. Normocephalic children, >6 months old, were selected for longitudinal follow-up using the Bayley Scales of Infant and Toddler Development (BSID-III) and the Ages & Stages Questionnaires: Social-Emotional (ASQ:SE-2). RESULTS: One hundred fifty-two children were assessed; after exclusion, 60 were exposed and 72 were unexposed to ZIKV during pregnancy. Twenty children in the exposed group and 21 children in the unexposed group had a composite score <85 in any of the BSID-III domains. Although exposed children had lower cognitive and language scores, differences were not statistically significant. For ASQ:SE-2 assessment, there were not statistically significant differences between groups. CONCLUSIONS: This study found no statistically significant differences in the neurodevelopment of normocephalic children between in utero ZIKV exposed and unexposed. Nevertheless, long-term monitoring of children with in utero ZIKV exposure is warranted. IMPACT: This study found no statistically significant differences in the neurodevelopment in normocephalic children with in utero Zika virus exposure compared to unexposed children, although the exposed group showed lower cognitive and language scores that persisted after adjustment by maternal age and education and after excluding children born preterm and low birth weight from the analysis. Children with prenatal Zika virus exposure, including those normocephalic and have no evidence of abnormalities at birth, should be monitored for neurodevelopmental delays. Follow-up is important to be able to detect developmental abnormalities that might not be detected earlier in life.

Low Levels of Lifetime Pap Test Receipt Among Vulnerable Guatemalans.

Low and middle-income countries, such as Guatemala, shoulder a disproportionate share of cervical cancer, a preventable disease in high income countries. Tangible obstacles, such as lack of access to health care, cultural differences, and insufficient infrastructure, and facilitators, such as being Ladino, married, and educated, have been identified in the literature related to cervical cancer prevention. The aim of this survey was to explore barriers and facilitators to cervical cancer prevention, comparing rural Indigenous and urban Ladino populations. We surveyed 139 women in two health clinics. Participants answered questions about demographic information, cervical cancer knowledge, and health care behaviors. We analyzed survey data with four bivariate models. Our results suggest vulnerable populations, such as rural Indigenous women who are single, illiterate, and lack education, face higher cervical cancer risk. Partnerships should be formed with health promotors and lay midwives to educate and encourage vulnerable populations to prevent cervical cancer.

Pretest-post-test evaluation with lay midwives in remote Guatemala after educational activities about COVID-19.

INTRODUCTION: Guatemalan lay midwives are well-respected community leaders in a country that lacks the institutional capacity to meet healthcare needs related to pregnancy, newborns, and COVID-19. Thus, Guatemalan lay midwives, who attend the majority of births in their country and who attend most births at home, are in an optimal position to offer frontline support to pregnant women and newborns regarding the global pandemic. The primary objective of this program of study was to dispel myths about COVID-19 and to provide culturally relevant educational activities to low-literacy Guatemalan lay midwives about issues related to the virus, such as signs and symptoms, risks to the mother and fetus, which patients are most vulnerable, appropriate responses, benefits and side-effects of the vaccine, timing of the vaccine, how the virus interacts with breastfeeding, and breastfeeding recommendations. METHODS: In a partnership among the Guatemalan Ministry of Health, expert faculty at the University of Utah College of Nursing, and Madre y Niño, a non-profit organization from the US, evidence-based educational activities about COVID-19 were offered orally in the native language of participants. Two hundred and ten lay midwives attended educational sessions at 11 locations throughout the remote Peten department. Educational activities included repetition, storytelling, and role plays. A pretest-post-test evaluation of 10 questions with 24 correct answers was used to determine if the educational activities changed lay midwife knowledge about COVID-19. Participants were given essential birth supplies and laminated COVID Reminder Cards, which were designed to increase visual literacy, to encourage knowledge retention after the educational sessions. RESULTS: Participants showed a significant increase in knowledge scores (possible 0-24) from prescores 7.09 (standard deviation (SD)=3.06) to 15.20 (SD=4.61), Student's t-test p<0.001. In addition, a significantly higher proportion of participants mistakenly thought COVID-19 passed through breast milk on the pretest (70.1%) compared to post-test (8.4%) (McNemar test, p<0.001). Regarding breastfeeding, 12.6% of participants knew on the pretest that women with COVID-19 who breastfeed should wear a mask and wash their hands compared to 74.3% of participants who knew these recommendations on the post-test (Wilcoxon signed-rank test, p<0.001). Finally, 2% of participants knew on the pretest that pregnant women with COVID-19 should take a low-dose aspirin compared to 67% of participants on the post-test (χ² (1)=194.7, p<0.001). CONCLUSION: These essential primary care providers misunderstood many critical issues related to COVID-19, pregnancy, and newborns. Culturally relevant educational activities provided orally in the native language of participants dispelled myths about the virus and significantly improved lay midwife knowledge. Providing evidence-based educational activities in a culturally relevant format is critical to protecting remote, vulnerable populations, such as pregnant Guatemalan women and newborns, during a global pandemic.

Zika virus evolution and spread in the Americas.

Although the recent Zika virus (ZIKV) epidemic in the Americas and its link to birth defects have attracted a great deal of attention, much remains unknown about ZIKV disease epidemiology and ZIKV evolution, in part owing to a lack of genomic data. Here we address this gap in knowledge by using multiple sequencing approaches to generate 110 ZIKV genomes from clinical and mosquito samples from 10 countries and territories, greatly expanding the observed viral genetic diversity from this outbreak. We analysed the timing and patterns of introductions into distinct geographic regions; our phylogenetic evidence suggests rapid expansion of the outbreak in Brazil and multiple introductions of outbreak strains into Puerto Rico, Honduras, Colombia, other Caribbean islands, and the continental United States. We find that ZIKV circulated undetected in multiple regions for many months before the first locally transmitted cases were confirmed, highlighting the importance of surveillance of viral infections. We identify mutations with possible functional implications for ZIKV biology and pathogenesis, as well as those that might be relevant to the effectiveness of diagnostic tests.

Efficient genome editing by CRISPR-Mb3Cas12a in mice.

As an alternative and complementary approach to Cas9-based genome editing, Cas12a has not been widely used in mammalian cells largely due to its strict requirement for the TTTV protospacer adjacent motif (PAM) sequence. Here, we report that Mb3Cas12a (Moraxella bovoculi AAX11_00205) can efficiently edit the mouse genome based on the TTV PAM sequence with minimal numbers of large on-target deletions or insertions. When TTTV PAM sequence-targeting CRISPR (cr)RNAs of 23 nt spacers are used, >70% of the founders obtained are edited. Moreover, the use of Mb3Cas12a tagged to monomeric streptavidin (mSA) in conjunction with biotinylated DNA donor template leads to high knock-in efficiency in two-cell mouse embryos, with 40% of founders obtained containing the desired knock-in sequences.

A PCR-RFLP method for the simultaneous differentiation of three Entamoeba species.

Amoebiasis caused by Entamoeba histolytica continues to be one of the most common parasitic diseases in the developing world. Despite its relevance, due to the lack of accurate diagnostic methods, the true clinical and public health importance of this parasite remains uncertain. The aim of this study was to develop a new diagnostic tool to differentiate E.histolytica from the morphologically undistinguishable E.dispar and E.moshkovskii. We developed a specific, fast and simple PCR-RFLP method that was able to accurately differentiate experimentally-obtained restriction patterns from the three Entamoeba species. This new method could prove useful for clinical and epidemiological purposes.

Sexuality Education Websites for Adolescents: A Framework-Based Content Analysis.

The web has unique potential for adolescents seeking comprehensive sexual health information. As such, it is important to understand the nature, scope, and readability of the content and messaging provided by sexuality educational websites. We conducted a content analysis of 14 sexuality education websites for adolescents, based on the 7 essential components (sexual and reproductive health and HIV, relationships, sexual rights and sexual citizenship, pleasure, violence, diversity, and gender) of the International Planned Parenthood Framework for Comprehensive Sexuality Education. A majority of content across all sites focused on sexual and reproductive health and HIV, particularly pregnancy and STI prevention, and other information about STIs and HIV. No other topic comprised more than 10% of content coverage across a majority of sites. The authors found little discussion of gender issues, sexual rights, sexual diversity, or sexual violence. Most sites provided brief references to sexual pleasure, generally moderated with cautionary words. Language used implied a heterosexual female audience. Reading levels for most sites were above the 9th-grade level, with several at the college level. These findings have implications for enhancing online sexuality education and broadening the coverage of essential topics.

San Raymundo: Focus Group About Comadronas' Response to Obstetrical Emergencies in Urban Guatemala.

INTRODUCTION: Comadronas attend most births in rural and urban Guatemala where the maternal mortality rate (MMR) is highest in Latin America. Information has been published regarding rural comadronas' response to obstetrical emergencies. Understanding urban comadronas' response to obstetrical emergencies is essential to addressing Guatemala's MMR. METHODS: A total of 17 urban comadronas participated in one, 34-min focus group to share their knowledge, practices, and attitudes regarding obstetrical emergencies. We used the long table to analyze the content to develop a matrix of themes. RESULTS: Five themes emerged. Urban comadronas receive consistent training, have hospital transportation, and feel confident in their knowledge, but they lack equipment and feel hospital providers disrespect them. Still, the joy of attending births outweighs the challenges they face. DISCUSSION: Urban comadronas described a different experience of responding to obstetrical emergencies than rural comadronas. Distinct approaches are needed to provide culturally congruent support for urban and rural comadronas when responding to obstetrical emergencies.

Justified emendation for a species-group name of longhorned beetle (Coleoptera, Cerambycidae, Psyrassa).

N/A.

Delivering Acupuncture Therapy in an Interdisciplinary Global Health Setting in Guatemala: Pilot Study and Lessons Learned.

Objective: Pain, stress, and diabetes mellitus (DM) are common complaints for Guatemalans seeking health care. Because acupuncture therapy (AT) is a low-cost, effective treatment for these concerns, it was offered to Guatemalans during a health care mission as an adjunct to primary care. The purpose of this study was to gather feedback about providing AT in this context and to describe the lessons that were learned. Materials and Methods: A pretest single-arm exploratory study design was used, collecting demographic data and administering a pretest to patients referred for AT. Patients received AT according to presenting symptoms, except patients with DM, who were treated with a previously developed auricular AT protocol. After AT, all patients received adhesive seeds for self-administered auricular acupressure, along with a printed diagram, showing where and how to apply them, and a symptom-tracking form. Results: AT was given to 11 patients (1 had DM; 10 did not). Most were female (9; 82%), older (average 59.27 years; range: 40-81 years), and had little education (average 4 years; range: 0-12 years). Complaints were pain (11; 100%), insomnia (6; 55%), anxiety (4; 36%), depression (7; 64%), and stress (3; 27%). More than 50% had seen health care providers (6; 55%). One person had AT previously and 3 people requested more information about AT before receiving it. Conclusions: Reporting high symptom burdens, most patients were unfamiliar with AT. AT ceased when the licensed acupuncturist contracted COVID. Patients will be followed in 2023 and AT will be given to collect data on feasibility, satisfaction, and possible implementation.

Evaluation of Transplacental Antibody Transfer in Pregnant Women Immunized with Different SARS-CoV-2 Homologous or Heterologous Schemes.

There is scarce information related to transplacental antibody transfer against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with different homologous and heterologous vaccination schemes. This study aimed to correlate the magnitude of transplacental transfer anti-SARS-CoV-2 antibodies in different homologous and heterologous schemes. An observational cross-sectional study was developed to identify pregnant women vaccinated against SARS-CoV-2. They were questioned about their immunization status; blood samples from the mother, umbilical cord during labor, and the newborn 72 h after birth were taken to measure anti-S1 and anti-S2 specific IgG antibodies for SARS-CoV-2. We recruited 104 women with a median age of 29 (SD 1.17). We found antibodies in all newborns with vaccinated mothers. Homologous BNT162b2 mRNA regimen had the highest mean (SD) antibody titers (AU/mL) in maternal (994.93 (3.08), p = 0.039), umbilical cord (1316.43 (2.79), p = 0.016), and newborn (1192.02 (3.55), p = 0.020) blood. The generalized linear model showed a positive effect over antibodies with at least one dose in maternal (ß = -1.1, p = 0.002) and newborn (ß= -0.717, p = 0.044) blood, and with two doses (ß = -0.684, p = 0.026) in umbilical cord blood. In conclusion, antibodies were detected in all vaccinated women and their newborns. Transfer of antibodies was found from the first dose, and the levels increased with the number of vaccine doses. Vaccination should be encouraged in pregnant women with any available scheme.

Draft Genome Sequence of Medusavirus Stheno, Isolated from the Tatakai River of Uji, Japan.

"Medusaviridae" is a proposed family of large double-stranded DNA (dsDNA) viruses so far represented by a sole virus isolated from a hot spring. In the present study, we report the isolation and genome sequencing of a second member of this family, medusavirus stheno, discovered from a freshwater sample with an Acanthamoeba castellanii coculture.

Evaluation of ELISA-Based Multiplex Peptides for the Detection of Human Serum Antibodies Induced by Zika Virus Infection across Various Countries.

Zika virus (ZIKV) is a mosquito-borne Flavivirus with a positive-sense RNA genome, which are generally transmitted through the bite of an infected Aedes mosquito. ZIKV infections could be associated with neurological sequelae that, and otherwise produces similar clinical symptoms as other co-circulating pathogens. Past infection with one member of the Flavivirus genus often induces cross-reactive antibodies against other flaviruses. These attributes complicate the ability to differentially diagnose ZIKV infection from other endemic mosquito-borne viruses, making it both a public health issue as well as a diagnostic challenge. We report the results from serological analyses using arbovirus-specific peptides on 339 samples that were previously collected from 6 countries. Overall, we found that our multiplexed peptide-based ELISA was highly efficient for identifying ZIKV antibodies as early as 2 weeks post infection, and that it correlates with microneutralization, plaque reduction neutralization tests (PRNTs) and commercial tests for ZIKV in previously characterized samples. We observed that seropositivity varied by patient cohort, reflecting the sampling period in relation to the 2015-2016 ZIKV outbreak. This work evaluates the accuracy, specificity, and sensitivity of our peptide-based ELISA method for detecting ZIKV antibodies from geographically diverse regions. These findings can contribute to ongoing serological methods development and can be adapted for use in future studies.

Microcephaly Outcomes among Zika Virus-Infected Pregnant Women in Honduras.

The impact of Zika virus (ZIKV) infection on pregnancies shows regional variation emphasizing the importance of studies in different geographical areas. We conducted a prospective study in Tegucigalpa, Honduras, recruiting 668 pregnant women between July 20, 2016, and December 31, 2016. We performed Trioplex real-time reverse transcriptase-PCR (rRT-PCR) in 357 serum samples taken at the first prenatal visit. The presence of ZIKV was confirmed in seven pregnancies (7/357, 2.0%). Nine babies (1.6%) had microcephaly (head circumference more than two SDs below the mean), including two (0.3%) with severe microcephaly (head circumference [HC] more than three SDs below the mean). The mothers of both babies with severe microcephaly had evidence of ZIKV infection. A positive ZIKV Trioplex rRT-PCR was associated with a 33.3% (95% CI: 4.3-77.7%) risk of HC more than three SDs below the mean.

Senales de Peligo: Guatemalan Lay Midwives' Perceptions of Responding to Obstetrical Complications.

Introduction: The Guatemalan maternal mortality rate is among the highest in Latin Americans, and lay midwives (LMs) attend home births. The study's purpose was to explore LMs' knowledge of, attitudes toward, and practices regarding obstetrical complications and emergencies. Method: In the remote Peten region, a descriptive ethnographic study held focus group discussions before education programs to teach/boost LMs' awareness about obstetrical complications. The long-table approach of analyses developed a matrix of common themes. Results: Nearly 200 LMs participated in 11 groups. Six themes emerged: LMs attribute their knowledge to God, recognize signs of obstetrical danger, want more training and equipment, encounter resistance from a patriarchal culture, feel fear and despair in addressing obstetrical complications/emergencies, and experience arduous logistics in emergency hospital transfers. Discussion: Contrary to published literature, LMs were able to verbalize basic knowledge of obstetrical complications. Information learned can guide future, action-research studies to address the maternal mortality rate in resource-poor settings.

Development and Validation of a Rapid Lateral Flow E1/E2-Antigen Test and ELISA in Patients Infected with Emerging Asian Strain of Chikungunya Virus in the Americas.

Since its 2013 emergence in the Americas, Chikungunya virus (CHIKV) has posed a serious threat to public health. Early and accurate diagnosis of the disease, though currently lacking in clinics, is integral to enable timely care and epidemiological response. We developed a dual detection system: a CHIKV antigen E1/E2-based enzyme-linked immunosorbent assay (ELISA) and a lateral flow test using high-affinity anti-CHIKV antibodies. The ELISA was validated with 100 PCR-tested acute Chikungunya fever samples from Honduras. The assay had an overall sensitivity and specificity of 51% and 96.67%, respectively, with accuracy reaching 95.45% sensitivity and 92.03% specificity at a cycle threshold (Ct) cutoff of 22. As the Ct value decreased from 35 to 22, the ELISA sensitivity increased. We then developed and validated two lateral flow tests using independent antibody pairs. The sensitivity and specificity reached 100% for both lateral flow tests using 39 samples from Colombia and Honduras at Ct cutoffs of 20 and 27, respectively. For both lateral flow tests, sensitivity decreased as the Ct increased after 27. Because CHIKV E1/E2 are exposed in the virion surfaces in serum during the acute infection phase, these sensitive and specific assays demonstrate opportunities for early detection of this emerging human pathogen.

Assessment of intensive agriculture on water quality in the Culiacan River basin, Sinaloa, Mexico.

The percentage of agricultural land cover effect on water quality in Culiacan River basin is studied in this research. The basin contains only intensive cropland as primary economic activity with 60% of the total area. Mathematical relationships between percentages of cropland and total phosphorus (TP) and total nitrogen (TN) concentrations were established. Sampling sites in middle and lower basin and water quality information during 2013-2018 were considered, and percentages of cropland were obtained by geospatial methods including variable area buffers. During rainy season, coefficients of determination were less than 0.2, although quantified nutrient concentration was higher, related to point sources of pollution in the basin. During dry season, coefficients of determination were higher than 0.76 and 0.90 for TN and TP, respectively, with an exponential mathematical trend. Results suggest that intensive agriculture practices generate accelerated loss of soil consolidation, which is transported to water bodies. These soils are in continuous contact with fertilizers and pesticides, mostly organophosphates which have been transported by runoff and underground flows. Using the information generated will help to establish environmental management plans, and to improve environmental diagnosis and effect in countries where there is not enough historical cartographic information and/or water quality data.

Suspected appendicitis in children: diagnostic importance of normal abdominopelvic CT findings with nonvisualized appendix.

PURPOSE: To determine whether lack of visualization of the appendix on otherwise normal abdominopelvic computed tomographic (CT) images can help exclude appendicitis in the pediatric population. MATERIALS AND METHODS: The study was institutional review board approved and HIPAA compliant. One thousand one hundred thirty-nine children suspected of having appendicitis were referred for CT examination between July 2002 and December 2006. Exclusion criteria included CT diagnosis of appendicitis or other cause of symptoms and lack of clinical follow-up. Consensus review was performed by two pediatric radiologists to determine normal examinations, leaving a final study group (nonvisualized appendix) of 156 patients (mean age, 9.6 years; boys, 7.2 years; girls, 10.2 years) and a control group (visualized appendix) of 421 patients (mean age, 11.0 years; boys, 9.8 years; girls, 11.2 years). In the control group, there were 168 subjects with a partially visualized (PV) appendix and 253 with a fully visualized (FV) appendix. Pericecal fat was graded according to published criteria. Diagnosis was confirmed at surgery or clinical follow-up. Negative predictive values were calculated with 95% confidence intervals (CIs). RESULTS: There were three false-negative findings (study group, two; control group, one [FV]). The negative predictive value of a normal CT examination in pediatric patients with a nonvisualized appendix was 98.7% (95% CI: 95.5%, 99.8%); that with a visualized appendix, 99.8% (95% CI: 98.7%, 99.99%); that with a PV appendix, 100% (95% CI: 97.8%, 100%); and that with a FV appendix, 99.6% (95% CI: 97.8%, 99.99%). CONCLUSION: Pediatric abdominopelvic CT images with nonvisualized appendix have a high negative predictive value, without significant difference from cases with a PV or even FV appendix. The false-negative rate was similar to those reported in two adult series.

Two new species of Beraba Martins, 1997 and new geographical records of Eburiini (Coleoptera, Cerambycidae).

Two new species of Beraba from Colombia (Bolívar) are described: Berabaanae sp. n. and Berabaangeli sp. n. The most recent key to species of the genus was modified to incorporate the new species. The male of Berabalimpida Martins, 1997 is described and illustrated for the first time. Moreover, the geographical distribution of 12 species of Eburiini is expanded.

Capturing sequence diversity in metagenomes with comprehensive and scalable probe design.

Metagenomic sequencing has the potential to transform microbial detection and characterization, but new tools are needed to improve its sensitivity. Here we present CATCH, a computational method to enhance nucleic acid capture for enrichment of diverse microbial taxa. CATCH designs optimal probe sets, with a specified number of oligonucleotides, that achieve full coverage of, and scale well with, known sequence diversity. We focus on applying CATCH to capture viral genomes in complex metagenomic samples. We design, synthesize, and validate multiple probe sets, including one that targets the whole genomes of the 356 viral species known to infect humans. Capture with these probe sets enriches unique viral content on average 18-fold, allowing us to assemble genomes that could not be recovered without enrichment, and accurately preserves within-sample diversity. We also use these probe sets to recover genomes from the 2018 Lassa fever outbreak in Nigeria and to improve detection of uncharacterized viral infections in human and mosquito samples. The results demonstrate that CATCH enables more sensitive and cost-effective metagenomic sequencing.